产肠毒素大肠杆菌F5菌毛基因的克隆与表达  

Clone and expression of pili subunit gene from adhesion F5 of the enterotoxigenic Escherichia coli(ETEC)

作　　者：于迪[1] 付海兵[1] 丁琳[1] 师东方[1] 单安山[2] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]东北农业大学动物科学技术学院,哈尔滨150030

出　　处：《东北农业大学学报》2008年第6期98-101,共4页Journal of Northeast Agricultural University

基　　金：东北农业大学校长基金;黑龙江省“十五”重点攻关项目(GB05B501);黑龙江省博士后资助经费

摘　　要：应用PCR方法从产肠毒素性大肠杆菌(ETEC)中扩增出不含信号肽序列的F5菌毛抗原基因片段,克隆测序后将其连接到大肠杆菌表达载体pET-30a(+)中,转化工程菌BL21,经IPTG诱导表达。经SDS-PAGE和Western Blot分析,重组蛋白在BL21中得到表达并与F5菌毛单因子血清发生明显反应。A F5 fimbrial subunit gene without signal peptide sequence was amplified by PCR from enterotoxigenic E. coll. After cloning and sequencing, the fragment was inserted into an E. coli expression vector pET- 30a（＋）, then the recombinant plasmid was transferred into E. coli BL21, and E. coli BL21（＋F5） obtained. Cultured and induced with IPTG, the expression of target protein was determined by SDS-PAGE and Western Blot. It was found that the expressed proteins could obviously react with antiserum against the single factor of F5 pili subunit proteins.

关 键 词：产肠毒素性大肠杆菌 F5菌毛 基因克隆 蛋白表达 

分 类 号：S852.612[农业科学—基础兽医学] Q785[农业科学—兽医学]